Parp 1 pcdna3.1 overexpression vector

Cell line and cell treatment INS-1 cells were obtained from the National Infrastructure of Cell Line Resource (Beijing, China) and cultured in RPMI-1640 (Invitrogen, Carlsbad, CA, USA) added with L-glutamine, FBS (10%; Invitrogen), HEPES (10 mmol/L; Sigma, St. Louis, MO, USA), sodium pyruvate (1 mmol/L; Invitrogen), and βmercaptoethanol (50 μmol/L; Sigma-Aldrich, St. Louis, MI, USA) at 37℃ in 5% CO 2 .
For high-fat induction, cholesterol (Santa Cruz Biotechnology; Dallas, TX, USA) was dissolved in chloroform and diluted to 2.5, 5 and 10 mM/L with culture medium for 24h; For GLP-1 treatment, cells were treated with GLP-1 agonist 5nM/L exendin-4 (Ex-4) for 24h; For AMPK activation or inhibition, cells were pre-treated with 0.5 mmol/L Acadesine (AICAR) or 10 μM/L compound c for 4 h and then co-treated with Ex-4 for 24 h, respectively. Cell transfection PARP-1 knockdown was generated in target cells by the transfection of si-PARP-1 synthesized by GenePharma (Shanghai, China). PARP-1 overexpression was generated in target cells by the transfection of PARP-1-pcDNA3.1 overexpression vector by (GenePharma). The transfection into target cells was performed with the help of lipofectamine 3000 (Invitrogen). Twenty-four hours later, the transfected cells were harvested for different experiments.